Expression miniarrays and uses thereof

• US 7,033,761 B2
• Filed: 11/14/2001
• Issued: 04/25/2006
• Est. Priority Date: 11/14/2000
• Status: Expired due to Fees

• Current Assignee: Genetag Technology, Inc.
• Original Assignee: David A. Shafer
• Inventors: Shafer, David A.
• Primary Examiner(s): WESSENDORF, TERESA D
• Application Number:
  US09/992,516
• Publication Number:
  US 20020074342A1
• Time in Patent Office: 1,623 Days
• Field of Search: 435/6, 435/4, 435/287.2, 435/287.3, 435/DIG.17, 536/22.1
• US Class Current: 435/6.11
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00369 in multiple or parallel arrangements
• CPC Class Codes: B01J 2219/00385 Printing
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01L 2400/02 : Drop detachment mechanisms of single droplets from nozzles or pins
• CPC Class Codes: B01L 2400/022 : droplet contacts the surface of the receptacle
• CPC Class Codes: B01L 2400/027 : electrostatic forces between substrate and tip
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 3/0262 : using touch-off at substrate or container
• CPC Class Codes: B01L 3/0268 : using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• CPC Class Codes: B01L 3/0275 : Interchangeable or disposable dispensing tips
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2035/00237 : Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 2035/103 : using disposable tips
• CPC Class Codes: G01N 2035/1034 : Transferring microquantities of liquid
• CPC Class Codes: G01N 33/54386 : Analytical elements
• CPC Class Codes: G01N 35/1002 : Reagent dispensers
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices
• CPC Class Codes: G01N 35/1074 : arranged in a two-dimensional array

Abstract

The present invention provides methods and devices for making new and inexpensive miniarrays suitable for gene expression analysis. Also provided herein are methods of diagnosis for specific tissue or condition using specialized diagnostic miniarrays that exhibit specific visual pattern as diagnostic readout.

Claims

1. A method for forming a miniarray on a miniarray substrate, said miniarray comprising locations or spots that contain an analyte specific reagent for detecting an analyte in a sample, said method comprises the steps of:

(a) aspirating a solution of each analyte specific reagent into dispensers comprising pipette tips spaced 9 mm or 4.5 mm center to center to load multiple reagent samples and said dispensers connected to a syringe pump;

(b) pressuring nanoliter quantities of defined droplets of said analyte specific reagent from the pipette tips of said dispensers that has been loaded with microliter quantities of said analyte specific reagent;

(c) touching said droplets to said miniarray substrate and causing said droplets to release through gravity which is perfomed by applying an electromechanical force to the tips of said dispensers, thereby spotting specific locations or spots on said substrate with a specific volume of said analyte specific reagent, wherein said locations or spots have a center-to-center spacing of in the range of 1 mm to 3 mm; and

(d) repeating steps (a) to (c) until said miniarray is fabricated.

2. The method of claim 1, wherein said dispensers are arranged in one or two rows.

3. The method of claim 1, wherein said dispensers have disposable tips that can be ejected and replaced automatically or fixed tips that are cleaned and dried between sample loadings.

4. The method of claim 1, wherein causing said droplet to release through gravity is performed by increasing pressure on said defined droplet to eject sufficient volume from the tips of said dispensers to cause said droplet to release by gravity.

5. The method of claim 1, wherein causing said droplet to release though gravity is performed by applying an electromechanical force to the tips of said dispensers to cause said droplet to release by gravity, wherein said electromechanical force is selected from the group consisting of vibration, piezoelectric pressure, and rapid mechanical actuation.

6. The method of claim 1, wherein said dispensers are carried by a robotically controlled apparatus that provides lateral and vertical motions, thereby automating the loading of multiple reagent samples, the replacement or cleaning of the pipette tips, and the spotting of multiple miniarrays under programmed instructions.

7. The method of claim 1, wherein said miniarray achieves a smaller, more condensed distribution by interspersing successive dispensing of reagents onto the array in regions between the spots dispensed previously.

8. The method of claim 1, wherein said dispensers are stationary, except for vertical motion, and miniarray substrates and reagent samples are moved under said dispensers by a robotic apparatus.

9. The method of claim 1, wherein said miniarray substrate is selected from the group consisting of coated microscope slides, flexible membranes, rigid glass, plastics, semi-rigid film, paper-based printing substrates, semi-rigid printing materials, photographic paper and high quality computer printing papers.